Genetic identification of degraded DNA samples buried in different types of soil

Biological samples buried in different types of soil are often found in crime scenes. These samples are usually highly degraded which difficult their analysis. Several factors contribute to the degradation of biological material including temperature variation, humidity, UV light and especially the presence of microorganisms.Blood was collected from three non-related male donors and blood stains were made in fabrics such as jeans, cotton and lycra. Blood stains were dried at room temperature and buried in three different types of soil (sand, marsh and clay), to promote its natural degradation.The buried samples suffered a high degradation over time which difficult their genetic identification. The marshy soil proved to be the most destructive one, leading to rapid degradation of the different analyzed fabrics, which disabled the obtainment of the genetic profiles.     

Y chromosome SNP analysis in a Portuguese population sample using a multiplex PCR and minisequencing strategy

The study of single nucleotide polymorphisms (SNPs) located on the Y chromosome specific region Y-SNPs (92R7, M70, M22, Tat, P25, SRY10831, M173, M213 and M9) was used to characterize a population sample from Central Portugal, in order to investigate the frequency distribution of the male lineages and to compare the observed results with those obtained in other Portuguese regions.The genotyping strategy was according to the described by Brion et al. [M. Brion, et al., Int. J. Legal Med. 119 (2004) 10-15].In this population sample from Central Portugal a typical Western European haplogroup composition was found. The majority of samples (almost 70%) were assigned inside haplogroup R. As for other Iberian populations, the most frequent haplogroup was R1b-P25 (52.2%), followed by F(xK)-M213 (15.2%), E-B-SRY10831.1, R1(xR1a,b)-M173 and R1a-SRY10831.2 (each of them with a 8.7% frequency), K2-M70 (4.3%) and L-M22 (2.2%). When comparing our sample with other samples from Portugal, no significant differences were found.     

The Confessional State in International Politics: Tudor England,Religion, and the Eclipse of Dynasticism

Whilst religion and collective identity have become issues of central concern to international relations scholars, dynastic concerns and national interests still dominate their analyses of early modern international politics. This analysis contributes to the constructivist emphasis on collective identity to foreign policy by examining Tudor England in light of the concept of confessionalisation. Based on the recent historiography of Tudor England, this analysis demonstrates the importance of religion in defining not only the collective identity of international actors, but also their foreign policies, choice of alliances, and, more generally, their international outlook. Through such a lens, it seeks to draw analyses of the confessional state away from their focus on domestic state formation to the “external” dimension of confessionalisation and its importance for international politics.     

‘Bringing the state back in’: A political economy analysis of Portuguese immigration policy

Southern European states are recurrently represented as ‘safe havens’ for irregular immigration in face of powerless national governments. Drawing from an interdisciplinary approach combining the domestic politics approach and the political economy of migration, this investigation explores the effectiveness of the Portuguese policy towards labour and irregular inflows during the 1990s and the 2000s. Against the increasing exclusion of politics and national governments in immigration studies, this research focuses on the agency of the Portuguese state. This article associates the intensity of irregular inflows with the covert expansive approach towards immigration adopted by the Portuguese state, driven by economic objectives.     

A study on ten short tandem repeat systems: African immigrant and Spanish population data

This work presents the results obtained from a genetic-population study for the D1S1656 system in the population of Southwest Spain (Huelva, Cádiz and Sevilla), Spaniards of Caucasian origin from North Africa (Ceuta), as well as in the black Central West African and Moroccan immigrant populations in Spain. The results of a study of the autochtonous population of the Canary Islands (n=138), and immigrant Central West African populations in Spain (n=132), obtained for nine short tandem repeat (STR) loci (D3S1358, VWA, FGA, D8S1179, D21S11, D18S51, D5S818, D13S317, D7S820), as well as the amelogenin locus, all contained in Profiler Plus (Perkin-Elmer) PCR amplification kits, are also presented. Except for the FGA and VWA data on immigrant Central West African populations in Spain, no deviations from the Hardy-Weinberg equilibrium were detected.     

The evolution of Japan's role in ‘Lusophone’ Africa: From inertia to action

This article explores Japan's relations with Lusophone Africa over the past 60 years. It asks what factors have propelled Japan's shift from a foreign policy based on inertia to a more proactive one. Arguably, colonialism and postcolonialism linked to the Cold War politics had a negative impact on Japan's relations with the Portuguese-speaking African countries (PALOP) and Africa as a whole; this contrasts with Japan's process of ‘Africanisation’ after 1990, reflective of changing external and internal circumstances. The question arises as to whether Japan's relations with the specific PALOP states, and with the group as a whole, have been influenced by, or have had influence over, Japan's long-term approach to other African states. Lastly, what characterises Japan's approach to the PALOP? Findings show mutual gains beyond the simple exploitation of natural resources towards broad-based sustainable growth. Still, from the perspective of the poorest PALOP, development cooperation and trade benefits are unbalanced and insufficient.     

Analysis of the Effect of Cyclophosphamide and Methotrexate on Chrysomya megacephala (Diptera: Calliphoridae),

Forensic entomotoxicology investigates the effects of chemical substances in the development of scavenger insects and the reflection on estimating the minimum postmortem interval (PMI). To evaluate the impact of the chemotherapeutic drugs cyclophosphamide (CF) and methotrexate (MTX) on the postembryonic development of Chrysomya megacephala, controlled experiments were performed by rearing the larvae on spiked minced beef with different concentrations of these drugs. The results indicated that CF significantly decreased their developmental rate up to 28 h but had no significant effect on larval and adult sizes, survival rate, and sex ratio, whereas MTX decreased larval and adult sizes, survival rate, and there was a deviation in the expected sex ratio toward females in MTX‐exposed larvae but had no significant impact on developmental rate. These negative interference factors should be considered in cases of suspected death of people that have undergone chemotherapy.     

A new technology in mtDNA sequencing: Success rates vs time

Study of mitochondrial DNA (mtDNA) control region is a current practice in forensic genetics. In our service, mtDNA analysis is performed in many evidentiary specimens. Evaluation of this methodology is important to improve quality, increase efficiency and decrease artefacts, in order to reduce costs and time consuming.A case with 12 reference samples (bucal swabs) and 190 telogenic hair specimens extracted with DNA IQ™ System Tissue and Hair Extraction Kit (Promega) is reported. HVS-1 and HVS-2 control regions were sequenced with BigDye^® Terminator v1.1 Kit (Applied Biosystems), using BetterBuffer (Microzone Limited), followed by a simple bead purification method (XTerminator) to remove unincorporated terminators. Application of this procedure had success in 180 hair samples within a very short time comparing to dRhodamine/ethanolic precipitation sequencing strategy and also demonstrated that better results are achieved with clean sequence data closer to the primer.The quality of data produced by the BigDye/BetterBuffer/XTerminator (BDX) procedure has been demonstrated to be very high. Besides that the BDX procedure can significantly reduce overall processing time and cost per reaction. This new methodology has additional advantages like fewer reagent transfers and smaller amounts of DNA.     

A DNA-based approach for the forensic identification of Asiatic black bear (Ursus thibetanus) in a traditional Asian medicine

Attempts to prevent illegal trade in bile and gallbladders from Asiatic black bears, Ursus thibetanus, are hampered by difficulties associated with identifying such items. We extracted DNA from bile crystals of unknown species origin and generated partial cytochrome b (cyt b) sequences using either universal primers (positioned in conserved regions of cyt b), or primers designed on existing U. thibetanus sequences (UT). Species origin was determined by aligning resolved sequences to reference sequence data. The universal primers were unsuitable for U. thibetanus identification when multiple species templates were present in the samples. The UT primers amplified U. thibetanus DNA from all sample extracts, including those containing mixed species templates. The amplified fragment can distinguish U. thibetanus from the most closely related species, U. americanus, a distinct advantage of DNA sequencing over the methods currently used to analyze suspected U. thibetanus bile.